[Usefulness of continuous glucose monitoring system (CGMS) in monitoring glycaemic profile in small children with diabetes type 1].
Improved methods of diabetes therapy result in a near normoglycaemic state in many patients. This leads however unfortunately to more frequent hypoglycaemic incidents. Particularly small children, whose nervous system is not fully mature, are at high risk of central nervous system damage in case of hypoglycaemia. A new method of detail monitoring of glycaemia provides CGMS system. The aim of the study was to compare the glycaemic profile, with high attention to hypoglycaemia in groups of young and older children with diabetes type 1, using CGMS and routine glucose meter. We studied 32 children with diabetes type 1. Children were divided into groups: group I--small children, n=17 (<7 yrs of age), mean age 5,8 years, with disease duration--2,46 years, with mean HbA1c level--7,22%, and group II--older children, n=15 (>10 years of age), mean age--12 years, with disease duration--3 years, with HbA1c level--7,21%. Continuous glucose monitoring system (CGMS), by MiniMed, was applied in outpatient or hospital conditions, after short training of patient and parents; together with routine glucose meter measurements, 4-8 times/24 hours. In 9 patients from small children group CGMS was repeated after 2 months. Hypoglycaemic incidents detected with CGMS were similar in both groups: 4,6 in I group vs. 4,2 in II group (ns). Hypoglycaemic incidents found with meter were lower in I group--1,6 vs. 2,3 in II group (ns). Mean hypoglycaemic time/24 hour was longer in small children group: 101 min vs. 74 min in group II (p<00,05). In I group we found higher number of hypoglycaemic incidents during the night compared to group II--1,7 vs. 0,8 (p<00,05) and longer duration of night hypoglycaemia: in I group--56 min vs. 32 min in group II (p<00,05). Repeated CGMS study in 9 children from I group revealed decreased mean time of hypoglycaemia/24 hours from 134 min/24 h to 90 min/24 h (p<00,05) and decreased time of night hypoglycaemia from 65 min to 40 min (p<00,05), with a comparable number of hypoglycaemic incidents. Hypoglycaemic incidents found with routine meter measurements in small children were 1,6 vs. 4,6 hypoglycaemia found with CGMS (p<00,05), in the older children group routine measurement found 2,3 hypoglycaemia vs. 4,2 detected with CGMS (ns). 1. CGMS can be particularly usefull in monitoring glucose profile and detecting hypoglycaemia incidents, mainly nocturnal in small children. 2. CGMS allows to verify meal dose of insulin and to decrease postprandial hyperglycaemia. 3. Modification of insulin therapy on the base of CGMS helps to decrease the time of hypoglycaemia and hyperglycemia, particularly during the night.


[Ferroptosis is involved in testicular injury induced by TDCIPP in adolescent male mice].
Objective: To investigate the role of ferroptosis in testicular injury in adolescent male mice induced by TDCIPP. Methods: In December 2021, 30 healthy 3-week-old male C57BL/6 mice, with a body weight of (13±2) g, were selected and fed adaptive for one week. They were divided into control group, low-dose group, medium-dose group, high-dose group and iron death inhibitor group according to a random number table, with 6 mice in each group. Mice in low, medium and high dose groups were treated with 5, 25 and 125 mg/ (kg·d) TDCIPP for 28 days, respectively, while the control group was treated with the same amount of corn oil for 28 days. The iron death inhibitor group was given 125 mg/ (kg·d) TDCIPP intragastric administration for 28 days, and 30 mg/kg DFO saline solution was intraperitoneally injected three times a week. After the treatment, the mice were killed, the epididymis was separated, and sperm count was performed. HE staining was used to observe the morphological changes of mouse testis, and iron content in testis was detected by tissue iron detection kit. The level of reactive cxygen species, MDA content, and the mitochondrial membrane potential level of mice were detected. Western blot analysis of testicular glutathione peroxidase (GPX4) and internal cyclooxygenase-2 (COX2) protein expression. Results: Compared with the control group, the spermatogenic cells in the testes of mice treated with medium-and high-dose of TDCIPP were disorderly arranged, showing a vacuolar structure. the number of sperm in the epididymis was significantly reduced (P=0.009, 0.004), while the sperm deformity rate was significantly increased (P=0.010, 0.000). Moreover, the content of ROS, iron ion and MDA in the testes increased significantly (P<0.05), and the mitochondrial membrane potential of mouse testicular cells decreased significantly (P<0.05). The expression of GPX4 proteins decreased (P<0.05). while the expression of COX2 increased significantly (P<0.01). Compared with high-dose group group, spermatogenic cells in ferroptosis inhibitor group were closely arranged and normal, and ROS and Fe contents in testicular tissue were significantly decreased (P<0.01) ; GPX4 protein expression was significantly increased while COX2 protein expression was significantly decreased (P<0.05) . Conclusion: Ferroptosis is involved in TDCIPP-induced testicular damage in male pubertal mice.


Real-Time Ultrasound-Guided Venous Access of the Arm for Right Heart Catheterization.
Arterial access from the wrist for cardiac catheterization is increasingly being used. Right heart catheterization (RHC) is an integral part of many of these procedures. Reliable venous access from the arm allows avoidance of femoral or jugular venous access for RHC. It is uncertain if ultrasound guidance offers a benefit for venous access of the arm for RHC. This study sought to assess the efficacy of ultrasound-guided venous access of the arm (UGVAA) for RHC. A retrospective study was performed on consecutive patients undergoing RHC at a single institution between August 2015 and July 2016. Baseline data, procedural information, and success rates of UGVAA and RHC were assessed. A total of 266 consecutive RHC procedures were identified, of which 253 (95.1%) were performed via arm venous access; of these, a pre-existing intravenous catheter was used in 3 cases, UGVAA was used in 241 cases, and UGVAA was probably used but not documented in 9 cases. There was 100% success of venous cannulation and sheath placement in these 253 patients. RHC via the arm vein was successful in 248 patients (98.0%) and failed in 5 patients (2.0%). Mean procedure time for RHC via arm access was 5.7 minutes. The femoral approach was used in 12 patients (4.5%). A jugular approach was used in 1 patient (0.4%). All patients had concomitant left heart catheterization via transradial access. UGVAA is a highly efficacious and safe technique, with a success rate of 98% for RHC in our consecutive series of 253 patients. UGVAA may allow for near-universal use of arm veins for RHC.


The emerging safety profile of JAK inhibitors in rheumatic disease.
Tofacitinib is the first Janus kinase (JAK) inhibitor commercially approved for the treatment of rheumatoid arthritis. This compound and a number of other JAK inhibitors are currently being tested in phase II and III trials for the treatment of a variety of autoimmune inflammatory diseases. Whereas a characteristic safety profile is emerging for some JAK inhibitors, differences between individual agents might emerge on the basis of distinct potency against their molecular targets. Similarly to biological therapy, JAK inhibition can lead to serious and opportunistic infections, and viral infections seem to be particularly frequent. Although no malignancy signals have been identified to date, long-term follow-up and further research are needed to understand the risk of malignancy associated with these compounds. As is the case for biologic agents, vaccination is important to mitigate the risks of these emerging therapies.


Effects of diaphragmatic contraction on lower limb venous return and central hemodynamic parameters contrasting healthy subjects versus heart failure patients at rest and during exercise.
The main objective was to assess the effects of abdominal breathing (AB) versus subject's own breathing on femoral venous blood flow (Qfv) and their repercussions on central hemodynamics at rest and during exercise contrasting healthy subjects versus heart failure (HF) patients. We measured esophageal and gastric pressure (PGA), Qfv and parameters of central hemodynamics in eight healthy subjects and nine HF patients, under four conditions: subject's own breathing and AB ( ∆: PGA ≥ 6 cmH2O) at rest and during knee extension exercises (15% of 1 repetition maximum) until exhaustion. Qfv and parameters of central hemodynamics [stroke volume (SV), cardiac output (CO)] were measured using Doppler ultrasound and impedance cardiography, respectively. At rest, healthy subjects Qfv, SV, and CO were higher during AB than subject's breathing (0.11 ± 0.02 vs. 0.06 ± 0.00 L·min(-1), 58.7 ± 3.4 vs. 50.1 ± 4.1 mL and 4.4 ± 0.2 vs. 3.8 ± 0.1 L·min(-1), respectively, P ≤ 0.05). ∆SV correlated with ∆PGA during AB (r = 0.89, P ≤ 0.05). This same pattern of findings induced by AB was observed during exercise (SV: 71.1 ± 4.1 vs. 65.5 ± 4.1 mL and CO: 6.3 ± 0.4 vs. 5.2 ± 0.4 L·min(-1); P ≤ 0.05); however, Qfv did not reach statistical significance. The HF group tended to increase their Qfv during AB (0.09 ± 0.01 vs. 0.07 ± 0.03 L·min(-1), P = 0.09). On the other hand, unlike the healthy subjects, AB did not improve SV or CO neither at rest nor during exercise (P > 0.05). In healthy subjects, abdominal pump modulated venous return improved SV and CO at rest and during exercise. In HF patients, with elevated right atrial and vena caval system pressures, these findings were not observed.


Delivery mode and intraventricular hemorrhage risk in very-low-birth-weight infants: Observational data of the German Neonatal Network.
Very-low-birth-weight infants (VLBWI) are frequently delivered by cesarean section (CS). However, it is unclear at what gestational age the benefits of spontaneous delivery outweigh the perinatal risks, i.e. intraventricular hemorrhage (IVH) or death. To assess the short-term outcome of VLBWI on IVH according to mode of delivery in a population-based cohort of the German Neonatal Network (GNN). A total cohort of 2203 singleton VLBWI with a birth weight <1500g and gestational age between 22 0/7 and 36 6/7 weeks born and discharged between 1st of January 2009 and 31st of December 2015 was available for analysis. VLBWI were stratified into three categories according to mode of delivery: (1) planned cesarean section (n=1381), (2) vaginal delivery (n=632) and (3) emergency cesarean section (n=190). Outcome was assessed in univariate and logistic regression analyses. Prevalence of IVH was significantly higher in the vaginal delivery (VD) (26.6%) and emergency CS group (31.1%) as compared to planned CS (17.2%), respectively. In a logistic regression analysis including known risk factors for IVH, vaginal delivery (OR 1.725 [1.325-2.202], p≤0.001) and emergency cesarean section (OR 1.916 [1.338-2.746], p≤0.001) were independently associated with IVH risk. In the subgroup of infants >30 weeks of gestation prevalence for IVH was not significantly different in VD and planned CS (5.3% vs. 4.4%). Our observational data demonstrate that elective cesarean section is associated with a reduced risk of IVH in preterm infants <30 weeks gestational age when presenting with preterm labor.


Interlaboratory study of cellular fluorescence intensity measurements with fluorescein-labeled microbead standards.
To determine the precision of cellular fluorescence intensity (FI) measurements derived from labeled microbead standards, FI results were compared from 43 different flow cytometers in 34 laboratories. All laboratories analyzed prepared aliquots of fluoresceinated calf thymocyte nuclei (Fluorotrol), human lymphocytes stained with fluoresceinated anti-CD4 antibody, and fluoresceinated microbeads used as both internal and external standards. Measurements were conducted by most laboratories on the third and fourth days after sample preparation. Results for percent of events within the gates and the histograms returned by participants indicated that the samples had remained stable and that gated populations had been properly identified. All standard curves showed strong linearity, and the pooled results from all standards produced a best-fit curve that was in close agreement with the assigned values. Nonetheless, results for cellular FI were highly variable, with CVs of 20-34%. Agreement within lab/instrument was much better, with CVs ranging from 3.0 to 9.9%. The overall variability was not obviously attributable to differences in the types of cytometer, nor could it be explained by attributes of the standard curves or any other single variable examined. However, the application of a corrective factor based on FI results for Fluorotrol allowed a two-fold improvement in the precision of FI measurements on CD4-stained lymphocytes, with an overall CV of 11%. Uncharacterized differences in the operating conditions of flow cytometers can influence cellular FI measurements, but consistent results can be obtained if a stained cellular calibrator is analyzed in addition to the proper microbead standards.


[Clinical aspect of new international gastric cancer staging system].
The 7th UICC/AJCC Gastric Cancer TNM Staging System includes major revisions of pT and pN classification. The Japanese Classification and UICC/AJCC TNM System have reached consistency in staging of gastric cancer. There are some new topics of lymphadenectomy in the new guidelines. The new TNM system accepts the database from Japan and Korea and it will be more accurate to predict the prognosis of gastric cancer patients. The rationality of splenectomy, total bursectomy, dissection of No.13 and No.14 lymph nodes is still not very clear and needs more evidences. D2 lymphadenectomy is the recommended surgical approach both in Eastern and Western countries. The benefit of paraaortic lymphadenectmoy for selected patients needs further evidences as well. The international gastric cancer staging project will collect the data from 23 countries and the new staging system will be applicable worldwide.


How can a binary switch within the pars tuberalis control seasonal timing of reproduction?
Life in seasonally changing environments is challenging. Biological systems have to not only respond directly to the environment, but also schedule life history events in anticipation of seasonal changes. The cellular and molecular basis of how these events are scheduled is unknown. Cellular decision-making processes in response to signals above certain thresholds regularly occur i.e. cellular fate determination, apoptosis and firing of action potentials. Binary switches, the result of cellular decision-making processes, are defined as a change in phenotype between two stable states. A recent study presents evidence of a binary switch operating in the pars tuberalis (PT) of the pituitary, seemingly timing seasonal reproduction in sheep. Though, how a binary switch would allow for anticipation of seasonal environmental changes, not just direct responsiveness, is unclear. The purpose of this review is to assess the evidence for a binary switching mechanism timing seasonal reproduction and to hypothesize how a binary switch would allow biological processes to be timed over weeks to years. I draw parallels with mechanisms used in development, cell fate determination and seasonal timing in plants. I propose that the adult PT is a plastic tissue, showing a seasonal cycle of cellular differentiation, and that the underlying processes are likely to be epigenetic. Therefore, considering the mechanisms behind adult cellular plasticity offers a framework to hypothesize how a long-term timer functions within the PT.


Stability of vertical posture explored with unexpected mechanical perturbations: synergy indices and motor equivalence.
We explored the relations between indices of mechanical stability of vertical posture and synergy indices under unexpected perturbations. The main hypotheses predicted higher posture-stabilizing synergy indices and higher mechanical indices of center of pressure stability during perturbations perceived by subjects as less challenging. Healthy subjects stood on a force platform and held in fully extended arms a bar attached to two loads acting downward and upward. One of the loads was unexpectedly released by the experimenter causing a postural perturbations. In different series, subjects either knew or did not know which of the two loads would be released. Forward perturbations were perceived as more challenging and accompanied by co-activation patterns among the main agonist-antagonist pairs. Backward perturbation led to reciprocal muscle activation patterns and was accompanied by indices of mechanical stability and of posture-stabilizing synergy which indicated higher stability. Changes in synergy indices were observed as early as 50-100 ms following the perturbation reflecting involuntary mechanisms. In contrast, predictability of perturbation direction had weak or no effect on mechanical and synergy indices of stability. These observations are interpreted within a hierarchical scheme of synergic control of motor tasks and a hypothesis on the control of movements with shifts of referent coordinates. The findings show direct correspondence between stability indices based on mechanics and on the analysis of multi-muscle synergies. They suggest that involuntary posture-stabilizing mechanisms show synergic organization. They also show that predictability of perturbation direction has strong effects on anticipatory postural adjustment but not corrective adjustments. We offer an interpretation of co-activation patterns that questions their contribution to postural stability.


The morphology of the right atrioventricular valve in the adult human heart.
Studies of the morphometry and normal anatomy of the tricuspid valve are in constant demand. Knowledge of the morphology of the normal tricuspid valve may be useful, for example in the context of the transfer of a leaflet of the tricuspid valve for repair or insufficiency of the mitral valve, in repair of the tricuspid valve after blunt chest trauma and in other surgical techniques of this region. In this study, performed in a group of 107 formalin-fixed adult human hearts, we attempted to assess the form and number of the main and accessory cusps in the tricuspid valve. Rare anatomical variants of the tricuspid valve were found. Using a planimeter we evaluated the surface area of the tricuspid valve and particular leaflets. With the help of a Vernier scale we measured the length and height of individual leaflets of the tricuspid valve and the length of the commissures. No differences were found between the length of the anterior and septal leaflets. The posterior leaflet was the shortest, while the anterior leaflet was the widest and had the largest surface area. The posterior leaflet was wider than the septal leaflet and had the smallest surface area. No differences were found between the main and accessory leaflets in the length of the commissures.


Convergent changes in melanocortin receptor 1 gene are associated with black-headed coat color in sheep.
As one of the most obvious phenotypic traits, the coat color of sheep is an ideal model to study the genetic mechanisms underlying coat color varieties of mammals. One distinguishable coat color is the black-headed type, such as the famous black-headed Dorper sheep from Africa and Bayinbuluke sheep from Asia. In this study, we compared the genome sequences of black-headed and all-white sheep to identify causative genes for the black-headed sheep, including black-headed Dorper vs. white-headed Dorper, as well as Bayinbuluke (black-headed) vs. Small-tailed Han (all-white). The most differentiating region between black-headed sheep and all-white sheep was found to harbor a haplotype covering melanocortin receptor 1 (MC1R) gene. The share of this haplotype by the black-headed sheep from Africa and Asia suggested that the convergent change in the MC1R region is likely to determine this unique coat color. Two missense mutations (g. 14251947T > A and g. 14252090G > A) within this haplotype of MC1R gene were found. We further analyzed whole genome sequence data of 460 worldwide sheep with diverse coat colors and confirmed the association between the MC1R haplotype with pigmentation variations. Our study provides novel insights into coat color genetics in sheep and expands our knowledge of the link between MC1R gene and varying pigmentation patterns in sheep.


Design and Synthesis of the Active Site Environment in Zeolite Catalysts for Selectively Manipulating Mechanistic Pathways.
By combining kinetics and theoretical calculations, we show here the benefits of going beyond the concept of static localized and defined active sites on solid catalysts, into a system that globally and dynamically considers the active site located in an environment that involves a scaffold structure particularly suited for a target reaction. We demonstrate that such a system is able to direct the reaction through a preferred mechanism when two of them are competing. This is illustrated here for an industrially relevant reaction, the diethylbenzene-benzene transalkylation. The zeolite catalyst (ITQ-27) optimizes location, density, and environment of acid sites to drive the reaction through the preselected and preferred diaryl-mediated mechanism, instead of the alkyl transfer pathway. This is achieved by minimizing the activation energy of the selected pathway through weak interactions, much in the way that it occurs in enzymatic catalysts. We show that ITQ-27 outperforms previously reported zeolites for the DEB-Bz transalkylation and, more specifically, industrially relevant zeolites such as faujasite, beta, and mordenite.


Anatomy of Escherichia coli sigma70 promoters.
Information theory was used to build a promoter model that accounts for the -10, the -35 and the uncertainty of the gap between them on a common scale. Helical face assignment indicated that base -7, rather than -11, of the -10 may be flipping to initiate transcription. We found that the sequence conservation of sigma70 binding sites is 6.5 +/- 0.1 bits. Some promoters lack a -35 region, but have a 6.7 +/- 0.2 bit extended -10, almost the same information as the bipartite promoter. These results and similarities between the contacts in the extended -10 binding and the -35 suggest that the flexible bipartite sigma factor evolved from a simpler polymerase. Binding predicted by the bipartite model is enriched around 35 bases upstream of the translational start. This distance is the smallest 5' mRNA leader necessary for ribosome binding, suggesting that selective pressure minimizes transcript length. The promoter model was combined with models of the transcription factors Fur and Lrp to locate new promoters, to quantify promoter strengths, and to predict activation and repression. Finally, the DNA-bending proteins Fis, H-NS and IHF frequently have sites within one DNA persistence length from the -35, so bending allows distal activators to reach the polymerase.


Does 6 Hours of Contact With Alginate Impression Material Affect Dental Cast Properties?
Alginate impression (irreversible hydrocolloid) material is commonly used in dental practice because it is easy to mix, low in cost, and well tolerated by patients. The material is not dimensionally stable, however; thus, it is necessary to pour the impression immediately after the molding is accomplished, or within 60 minutes if the impression is kept in 100% humidity. Excessive contact of the alginate impression with the cast model over time may affect the model's properties. In this study, the authors tested the effect of contact time between an alginate impression and type III dental stone on cast model properties. Sixty-seven cast models were obtained from a stainless steel cylinder by using irreversible hydrocolloid impression material and type III dental stone. Thirty-seven cast models were separated from the impression after 1 hour (control group) and 30 cast models were separated after 6 hours (study group). The samples were evaluated under light microscope for surface details and measured by digital caliper for dimensional stability. An indentation on the cast was made and the depth of the indentation was then measured with a digital caliper to measure hardness. The dimensional stability of the cast models was not affected when contact time was increased from 1 hour to 6 hours (P = .507). Surface details did not deteriorate when contact time was increased, as all of the samples could reproduce all details after the 1-hour and 6-hour interval periods. However, hardness was greater after 1 hour of contact time (P = .001) than after 6 hours of contact time. In conclusion, contact between alginate impression material and type III dental stone up to 6 hours did not affect the dimensional stability and richness of the surface; hardness, though, was significantly affected.


Use of Nanoindentation in Determination of Regional Biomechanical Properties of Rabbit Cornea After UVA Cross-Linking.
To evaluate the regional effects of different corneal cross-linking (CXL) protocols on corneal biomechanical properties. The study involved both eyes of 50 rabbits, and the left eyes were randomized to the five intervention groups, which included the standard CXL group (SCXL), which was exposed to 3-mW/cm2 irradiation, and three accelerated CXL groups (ACXL1-3), which were exposed to ultraviolet-A at irradiations of 9 mW/cm2, 18 mW/cm2, and 30 mW/cm2, respectively, but with the same total dose (5.4 J/cm2). A control (CO) group was not exposed to ultraviolet-A. No surgery was done on the contralateral eyes. The corneas of each group were evaluated by the effective elastic modulus (Eeff) and the hydraulic conductivity (K) within a 7.5-mm radius using nanoindentation measurements. Compared with the CO group, Eeff (in regions with radii of 0-1.5 mm, 1.5-3.0 mm, and 3.0-4.5 mm) significantly increased by 309%, 276%, and 226%, respectively, with SCXL; by 222%, 209%, and 173%, respectively, with ACXL1; by 111%, 109%, and 94%, respectively, with ACXL2; and by 59%, 41%, and 37%, respectively, with ACXL3 (all P < 0.05). K was also significantly reduced by 84%, 81%, and 78%, respectively, with SCXL; by 75%, 74%, and 70%, respectively, with ACXL1; by 64%, 62%, and 61%, respectively, with ACXL2; and by 33%, 36%, and 32%, respectively, with ACXL3 (all P < 0.05). For the other regions(with radii between 4.5 and 7.5 mm), the SCXL and ACXL1 groups (but not the ACXL2 and ACXL3 groups) still showed significant changes in Eeff and K. CXL had a significant effect on corneal biomechanics in both standard and accelerated procedures that may go beyond the irradiated area. The effect of CXL in stiffening the tissue and reducing permeability consistently decreased with reducing the irradiance duration.


Influence of carbohydrate and age on lymphocyte function following a marathon.
The influence of 6% carbohydrate ingestion and age on PHA-induced lymphocyte proliferation and in vitro cytokine production was studied in 48 runners following a competitive marathon. Runners were randomly assigned to carbohydrate (C; n = 23) and placebo (P; n= 25) groups, with blood samples taken before, immediately after, and 1.5 hr post-race. C versus P ingestion resulted in higher plasma glucose, lower plasma cortisol, reduced neutrophilia, and monocytosis during recovery, but had no effect on the post-exercise reduction in T-lymphocytes or NK cells, or on race times. No group differences were observed for PHA-induced lymphocyte proliferation or cytokine production. However, for all subjects combined, lymphocyte proliferation and IFN-gamma secretion decreased significantly below pre-race values by 1.5 hr of recovery, and these were negatively correlated with plasma cortisol. Young (<50 years; n = 36) and old (>or=50 years; n = 12) runners exhibited parallel post-race declines in lymphocyte proliferation and IFN-gamma secretion, with the older group exhibiting a 33-59% lower proliferation at each time point. In conclusion, PHA-induced lymphocyte proliferation and cytokine production decreased significantly following a marathon, and this decrease was strongly linked to cortisol and only partially linked to T-cell changes. This decrease occurred in both younger and older runners and was not influenced by carbohydrate.


The effect of 2-micron DNA on survival and mutagenesis in Saccharomyces cerevisiae.
Strains of Saccharomyces cerevisiae, with and without endogenous 2-microns DNA, were studied in experiments designed to determine the effect of this plasmid on survival and mutagenesis in yeast. Comparison of the two strains exposed to ultraviolet light, 4-nitroquinoline oxide, or methyl methanesulfonate (MMS), revealed that the presence of 2-microns DNA slightly enhanced survival after exposure to each agent. Spontaneous frequencies of mutations (histidine reversion, canavanine resistance, and mitochondrial petites, but not adenine auxotrophy) were reduced by the presence of 2-microns DNA. MMS-induced His+ reversion was weak, and both strains responded similarly. No difference was found between the two strains when induced forward mutation to canavanine resistance was examined. The extent of induction of mitochondrial petites was about the same in both strains. Therefore, it appears that under these experimental conditions with these mutagens, 2-microns DNA has an effect on spontaneous mutation and survival after DNA damage but not on induced mutagenesis in S. cerevisiae.


Computational methods for in situ structural studies with cryogenic electron tomography.
Cryo-electron tomography (cryo-ET) plays a critical role in imaging microorganisms in situ in terms of further analyzing the working mechanisms of viruses and drug exploitation, among others. A data processing workflow for cryo-ET has been developed to reconstruct three-dimensional density maps and further build atomic models from a tilt series of two-dimensional projections. Low signal-to-noise ratio (SNR) and missing wedge are two major factors that make the reconstruction procedure challenging. Because only few near-atomic resolution structures have been reconstructed in cryo-ET, there is still much room to design new approaches to improve universal reconstruction resolutions. This review summarizes classical mathematical models and deep learning methods among general reconstruction steps. Moreover, we also discuss current limitations and prospects. This review can provide software and methods for each step of the entire procedure from tilt series by cryo-ET to 3D atomic structures. In addition, it can also help more experts in various fields comprehend a recent research trend in cryo-ET. Furthermore, we hope that more researchers can collaborate in developing computational methods and mathematical models for high-resolution three-dimensional structures from cryo-ET datasets.


High-Quality Axions in a Class of Chiral U(1) Gauge Theories.
We show that there are many candidates for the quintessence and/or the QCD axions in a class of chiral U(1) gauge theories. Their qualities are high enough to serve as the dark energy and/or to solve the strong CP problem. Interestingly, the high quality of axion is guaranteed by the gauged U(1) and Z_{2N} symmetries and hence free from the nonperturbative quantum gravity corrections. Furthermore, our mechanism can be easily applied to the Fuzzy dark matter axion scenarios.


Proton therapy planning and image-guidance strategies within a randomized controlled trial for high-risk prostate cancer.
The Danish Prostate Cancer Group is launching the randomized trial, PROstate PROTON Trial 1 (NCT05350475), that compares photons and protons to the prostate and pelvic lymph nodes in treatment of high-risk prostate cancer. The aim of the work described in this paper was, in preparation of this trial, to establish a strategy for conventionally fractionated proton therapy of prostate and elective pelvic lymph nodes that is feasible and robust. Proton treatments are image-guided based on gold fiducial markers and on-board imaging systems in line with current practice. Our established proton beam configuration consists of four coplanar fields; two posterior oblique fields and two lateral oblique fields, chosen to minimize range uncertainties associated with penetrating a varying amount of material from both treatment couch and patient body. Proton plans are robustly optimized to ensure target coverage while keeping normal tissue doses as low as is reasonably achievable throughout the course of treatment. Specific focus is on dose to the bowel as a reduction in gastrointestinal toxicity is the primary endpoint of the trial. Strategies have been established using previously treated patients and will be further investigated and evaluated through the ongoing pilot phase of the trial.


Bone penetration of intravenous flucloxacillin and gentamicin as antibiotic prophylaxis during total hip and knee arthroplasty.
To investigate the bone penetration of intravenous antibiotic prophylaxis with flucloxacillin and gentamicin during hip and knee arthroplasty, and their efficacy against Staphylococcus (S.) aureus and S. epidermidis. Bone samples from the femoral head, neck and acetabulum were collected from 18 patients undergoing total hip arthroplasty (THA) and from the femur and tibia in 21 patients during total knee arthroplasty (TKA). The concentration of both antibiotics in the samples was analysed using high performance liquid chromatography. Penetration was expressed as a percentage of venous blood concentration. The efficacy against common infecting organisms was measured against both the minimum inhibitory concentration 50, and the more stringent epidemiological cutoff value for resistance (ECOFF). The bone penetration of gentamicin was higher than flucloxacillin. Relative to ECOFF, flucloxacillin concentrations were effective against S. aureus and S. epidermidis in all THAs and 20 (95%) TKAs. Gentamicin concentrations were effective against S. epidermidis in all bone samples. Gentamicin was effective against S. aureus in 11 (61.1%) femoral neck samples in THA. Effective concentrations of gentamicin against S. aureus were only achieved in four (19%) femoral and six (29%) tibial samples in TKA. Flucloxacillin and gentamicin were found to penetrate bone during THA and TKA. Gentamicin was effective against S. epidermidis in both THA and TKA, while levels were subtherapeutic against S. aureus in most TKAs. Bone penetration of both antibiotics was less in TKA than THA, and may relate to the use of a tourniquet. Using this antibiotic combination, effective cover against the two common infective organisms was achieved in all THAs and all but one TKA. Cite this article: Bone Joint J 2017;99-B:358-64.


[Prevalence of cryptosporidiosis in pediatric hospital patients in Niamey, Niger].
No data has been published, so far about the prevalence of cryptosporidiosis among children in Niger; a landlocked country of West Africa where malnutrition and diarrhoea are two major public health issues. The aim of the present study was to get a first evaluation of the prevalence of Cryptosporidium sp in the stools of hospitalized children in the National Hospital of Niamey (NHN) where we carried out a prospective descriptive study involving all children younger than 5 years of age in the paediatric department between February 21st and May 22nd, 2004. Direct stool examination and Ritchie technique were systematically performed, and evidence of Cryptosporidium oocysts was looked for by means of a smear from the sediment which was stained by the modified Ziehl-Neelsen technique. The weight/age ratio was calculated and analyzed with the Epi-Info software, based on the reference population defined by the US National Center for Health Statistics (NCHS). Malnutrition was defined as a weight/age ratio more than 2 SD below the NCHS's reference population. Malnutrition was considered moderate between -2 and -3 SD and severe below 3 SD. In the 3 months study 220 children were included (sex ratio = 1.18 and mean age = 20 months) showing that 65% of the children were suffering from malnutrition (moderate = 17.3%, severe = 47%). Diarrhoea was reported in 51.8% of the children. Cryptosporidium oocysts were detected in 12/220 children (5.5% of the studied population) and 7/114 (6. 1%) of those children were suffering from diarrhoea. 10 (83%) out of the 12 infected children were malnourished. This was the first study ever conducted in the paediatric department of the Niamey hospital, and it showed evidence of a 5.5% prevalence of cryptosporidiosis in the overall studied population, versus 6.1% among children with diarrhoea. 5 children without diarrhoea and 2 children under 6 months were also infected. Another study based on a larger number of patients would be necessary to address the impact of rainfall distribution on the incidence of the disease.


Evaluating patient-reported outcome measures (PROMs) for future clinical trials in adult patients with optic neuritis.
To search for and critically appraise the psychometric quality of patient-reported outcome measures (PROMs) developed or validated in optic neuritis, in order to support high-quality research and care. We systematically searched MEDLINE(Ovid), Embase(Ovid), PsycINFO(Ovid) and CINAHLPlus(EBSCO), and additional grey literature to November 2021, to identify PROM development or validation studies applicable to optic neuritis associated with any systemic or neurologic disease in adults. We included instruments developed using classic test theory or Rasch analysis approaches. We used established quality criteria to assess content development, validity, reliability, and responsiveness, grading multiple domains from A (high quality) to C (low quality). From 3142 screened abstracts we identified five PROM instruments potentially applicable to optic neuritis: three differing versions of the National Eye Institute (NEI)-Visual Function Questionnaire (VFQ): the 51-item VFQ; the 25-item VFQ and a 10-item neuro-ophthalmology supplement; and the Impact of Visual Impairment Scale (IVIS), a constituent of the Multiple Sclerosis Quality of Life Inventory (MSQLI) handbook, derived from the Functional Assessment of Multiple Sclerosis (FAMS). Psychometric appraisal revealed the NEI-VFQ-51 and 10-item neuro module had some relevant content development but weak psychometric development, and the FAMS had stronger psychometric development using Rasch Analysis, but was only somewhat relevant to optic neuritis. We identified no content or psychometric development for IVIS. There is unmet need for a PROM with strong content and psychometric development applicable to optic neuritis for use in virtual care pathways and clinical trials to support drug marketing authorisation.


Activities by the schools to improve the distribution of dentists: an AADS report.
It appears that most dental schools have not yet become actively involved in efforts to solve maldistribution problems. However, there seems to be increasing interest in the potential benefits of extramural experience programs. Coordination of efforts by the schools, the state dental associations, and the state boards of dental examiners is very limited, but is clearly needed. Admissions committees seem to be trying to take into account the needs of rural areas and the inner city as well. Finally, the results of the survey point up the need for further study of these problems and of the value of particular approaches to them.


Lentiviral correction of enzymatic activity restrains macrophage inflammation in adenosine deaminase 2 deficiency.
Adenosine deaminase 2 deficiency (DADA2) is a rare inherited disorder that is caused by autosomal recessive mutations in the ADA2 gene. Clinical manifestations include early-onset lacunar strokes, vasculitis/vasculopathy, systemic inflammation, immunodeficiency, and hematologic defects. Anti-tumor necrosis factor therapy reduces strokes and systemic inflammation. Allogeneic hematopoietic stem/progenitor cell (HSPC) transplantation can ameliorate most disease manifestations, but patients are at risk for complications. Autologous HSPC gene therapy may be an alternative curative option for patients with DADA2. We designed a lentiviral vector encoding ADA2 (LV-ADA2) to genetically correct HSPCs. Lentiviral transduction allowed efficient delivery of the functional ADA2 enzyme into HSPCs from healthy donors. Supranormal ADA2 expression in human and mouse HSPCs did not affect their multipotency and engraftment potential in vivo. The LV-ADA2 induced stable ADA2 expression and corrected the enzymatic defect in HSPCs derived from DADA2 patients. Patients' HSPCs re-expressing ADA2 retained their potential to differentiate into erythroid and myeloid cells. Delivery of ADA2 enzymatic activity in patients' macrophages led to a complete rescue of the exaggerated inflammatory cytokine production. Our data indicate that HSPCs ectopically expressing ADA2 retain their multipotent differentiation ability, leading to functional correction of macrophage defects. Altogether, these findings support the implementation of HSPC gene therapy for DADA2.


Influence of a novel platinum compound--cis-dichloro (dimethylsulphoxide) (1-beta-D-rybofuranosyl-1,2,4-triazolo-3-arboxyamide) platinum (II)--"Pt-rib-1"--on cell cycle and apoptosis in ClS91 and B16 mouse melanoma in vitro.
Cisplatin has a significant role in the treatment of selected human tumors including advanced melanoma, but new platinum compounds are still in focus of search for better properties. Modern drug design is often based on studies detecting the abilities of tested drug to induce apoptosis and disturb cell cycle. Aim of the study was to establish the influence of a platinum complex Pt-rib-1 on cell cycle and apoptosis occurrence in mouse melanoma B16 and ClS91 cells. Pt-rib-1 is a ribavirin derivative. previously characterized and described as cytotoxic to B16 and ClS91 mouse melanoma cells in vitro. The new platinum complex (Pt-rib-1); cis- dichloro (dimethylsulphoxide) (1- beta- D-ribofuranosyl- 1,2,4-triazolo -3- carboxyamide) platinum (II) was supplied. Cisdiaminodichloroplatinum (II), (cisplatin) was used in control groups. To detect apoptotic and necrotic cells, Annexin V- conjugated with fluorescein isothiocyanate (Annexin V-FITC, Immunotech) and propidium iodide (IP, Immunotech) were used. Apoptosis detection were done using fluorescence microscopy and flow cytometry. The total DNA content within the cell indicated phase of the cell cycle. DNA content was measured using flow cytometry. Values given represent the mean from three determinations. Results were presented as mean +/- standard deviation (SD). Statistical analysis was done using t-Student test. There were 70.4% of apoptotic cells in the ClS91 culture after 24 h incubation with Pt-rib-1 at a concentration of 2.04 x 10(-3) M (4 x IC50). In B16 group, 83.2 per cent of apoptotic cells was found after 24h incubation with Pt-rib-1 at high concentration (2.30 x 10(-3) M). A 24-h experiment shows a threshold at a concentration higher than 3 x IC50 responsible for apoptosis induction in B16 and ClS91 cells. After 48 h incubation with Pt-rib-1 the per cent of apoptotic cells increased gradually with rising concentrations of Pt-rib-1 up to a final concentration of 2.04 x 10(-3) M and 0.92 x 10(-3) M in ClS91 and B16 groups, respectively. Cell accumulation was observed in S phase after 48 h incubation with Pt-rib-1. The per cent of cells in S phase increased from 31 to 51.1% and 38.8 to 50.0% in ClS91 and B16 culture, respectively. There were no B16 and ClS91 cells in G2/M phase after incubation with higher concentrations of Pt-rib-1 (from 0.2 to 2.0 x 10(-5) M/dm3). Pt-rib-1 partially exhibits action of cisplatin. which has no specific influence on cell cycle and ribavirin. probably responsible for DNA synthesis delay.


[Prevention of infections in patients with inflammatory bowel disease during TNF inhibitor treatment].
Treatment of inflammatory bowel disease with anti-tumour necrosis factor (TNF)-alpha biopharmaceuticals is generally well tolerated, but increases the risk of infections due to globally induced suppression of the immune system. National and international guidelines recommend screening and vaccination for selected infectious agents prior to anti-TNF initiation as well as during ongoing anti-TNF therapy. This review focuses on current available recommendations and discusses physicians' and patients' barriers for adherence to guidelines.


A combined cumulative threshold spectra and digital reconstruction analysis reveal structural alterations of microglia within the prefrontal cortex following low-dose LPS administration.
Sickness behaviors have become the focus of great interest in recent years as they represent a clear case of how peripheral disturbances in immune signaling can disrupt quite complex behaviors. In the current study, we were interested in examining whether we could identify any significant morphological disturbances in microglia associated with these sickness-like behaviors in adult male Sprague-Dawley rats. We chose lipopolysaccharide (LPS 100 μg/kg/i.p.), to induce sickness-like behaviors as it is the most well-validated approach to do so in rodents and humans. We were particularly interested in examining changes in microglia within the prefrontal cortex (PFC) as several recent neuroimaging studies have highlighted significant functional changes in this region following peripheral LPS administration. Paraformaldehyde-fixed tissue was collected from animals 24h post LPS administration and labeled immunohistochemically with an antibody directed to bind to Iba-1, a protein known to be involved in the structural remodeling of microglia. To analyze changes, we have made use of two recently described image analysis procedures. The first is known as cumulative threshold spectra (CTS) analysis. The second involves the unsupervised digital reconstruction of microglia. We undertook these complementary analysis of microglial cells in the both the pre- and infralimbic divisions of the PFC. Our results indicated that microglial soma size was significantly enlarged, while cell processes had contracted slightly following LPS administration. To our knowledge this study is to first to definitely demonstrate substantial microglial disturbances within the PFC following LPS delivered at a dose that was sufficient to induce significant sickness-like behavior.


Historical contingency and the role of post-invasion evolution in alternative community states.
Historical contingency has long figured prominently in the conceptual frameworks of evolutionary biology and community ecology. Evolutionary biologists typically consider the effects of chance mutation and historical contingency in driving divergence and convergence of traits in populations, whereas ecologists instead are often interested in the role of historical contingency in community assembly and succession. Although genetic differences among individuals in populations can influence community interactions, variability among populations of the same species has received relatively little attention for its potential role in community assembly and succession. We used a community-level study of experimental evolution in two compositionally different assemblages of protists and rotifers to explore whether initial differences in species abundances among communities attributed to differences in evolutionary history, persisted as species that continued to evolve over time. In each assemblage, we observed significant convergence between two invaded treatments initially differing in evolutionary history over an observation period equal to ~40-80 generations for most species. Nonetheless, community structure failed to converge completely across all invaded treatments within an assemblage to a single structure. This suggests that whereas the species in the assemblage represent a common selective regime, differences in populations reflecting their evolutionary history can produce long-lasting transient alternative community states. In one assemblage, we also observed increasing within-treatment variability among replicate communities over time, suggesting that ecological drift may be another factor contributing to community change. Although subtle, these transient alternative states, in which communities differed in the abundance of interacting species, could nonetheless have important functional consequences, suggesting that the role of evolution in driving these states deserves greater attention.


Radial chorioretinal folds. A sign of choroidal neovascularization.
A radial pattern of chorioretinal folds surrounding the margins of a localized macular detachment occurred in four patients with drusen of Bruch's membrane. Biomicroscopic and angiographic findings suggested that contraction of a subpigment epithelial choroidal neovascular membrane caused a focal area of shrinkage, or puckering, of the choroid and a series of chorioretinal folds radiating outward from the edge of the membrane. In two eyes, these radial folds developed early in the course of the macular detachment before any other reliable biomicroscopic signs of choroidal neovascularization were present.


[Functional evaluation and quality of life in the follow-up of patients with Charcot arthropathy treated by ankle arthrodesis with a locked retrograde nail].
Charcot's arthropathy is a disabling non-infectious, progressive condition characterized by bony and articular destruction in patients with sensory neuropathy. In advanced cases with deformities and ankle instability, it requires a more invasive treatment as an ankle fusion with a retrograde locked intramedullary nail. It is unknown if, in these patients, the functional results of AOFAS (American Orthopaedic Foot and Ankle Society) correlate with the quality of life ones from the EuroQol-5D test and the possible postoperative complications. the design is experimental, longitudinal prospective with ambispective data analysis (retrospective and prospective) to evaluate the functional results and life quality with a year of following patients with Charcot's arthropathy diagnosis treated with a retrograde locked nail from January 1, 2010, to November 1, 2018. this study resulted in complete consolidation from nine out of 11 cases, with a success rate of 81.2%, and only two cases (18.2%) developed nonunion. AOFAS and EuroQol-5D tests correlate positively in agreement with the Pearson correlation. AOFAS and EuroQol-5D correlate positively, having a 45 and 63% of acceptable and satisfactory results, respectively, for both tests.


Ferredoxin reductase: pharmacogenomic assessment in colorectal cancer.
Ferredoxin reductase (FDXR) is a putative contributor to TP53-mediated apoptosis from 5-fluorouracil chemotherapy through the generation of oxidative stress. With TaqMan real-time quantitative reverse transcription-PCR, this study established a significant difference in FDXR relative RNA expression level between tumor (median, 212.9 units) and normal tissues (median, 113.8 units) from 51 colorectal cancer patients (P < 0.001). Seven single nucleotide polymorphisms (SNPs) in the FDXR gene were discovered, with no significant difference in variant allele frequency between colon tumor and normal tissues (P > 0.05), and the common haplotypes for FDXR were not different between colon tumor and normal samples. No correlation was observed between FDXR genotype and RNA expression implying that the polymorphisms described in this study are not regulating FDXR expression in colon cancer. This genomic characterization provides the foundation for pharmacogenetic analysis of the impact of FDXR on chemotherapy for colorectal cancer.


Selection of surgical or percutaneous coronary intervention provides differential longevity benefit.
Treatment of coronary artery disease (CAD) is evolving with better medications, improvements in percutaneous coronary intervention (PCI), and enhanced techniques for coronary artery bypass grafting (CABG). In this study, 18,481 patients with significant (>75% stenosis) CAD treated at a single center between 1986 and 2000 were assigned to one of three groups based on initial treatment strategy: medical therapy (MED) (n = 6862), PCI (n = 6292), or CABG (n = 5327). Each group was categorized into 3 groups according to baseline severity of CAD: low-severity (predominantly 1-vessel), intermediate-severity (predominantly 2-vessel), and high-severity (all 3-vessel), and prospectively evaluated in Cox models for all-cause mortality adjusted for cardiac risk, comorbidity, and propensity for selection of a specific treatment. Treatments were compared for the entire period and three eras (1: 1986 to 1990; 2: 1991 to 1995; 3: 1996 to 2000), the last encompassing widespread availability of PCI with stenting. Survival significantly improved in all groups for all degrees of CAD, despite increasing severity of illness. Revascularization strategies provided significant survival over MED with 8.1, 10.6, and 23.6 additional months per 15 years of follow-up for low-severity, intermediate-severity, and high-severity CAD, respectively. Therapeutic improvements led to increased survival of 5.3 additional months per 7 years of follow-up (95% confidence interval, 0.2 to 10.2; p = 0.039) in era 3 for CABG compared with PCI for high-severity CAD. Initial revascularization strategies result in significant survival advantage over MED for all CAD levels. Patients with high-severity CAD have reduced survival with PCI compared with those initially treated with CABG.


Cervical spondylotic myelopathy with syringomyelia presenting as hip Charcot neuroarthropathy: A case report and review of literature.
Charcot neuroarthropathy (CN) is a systemic disease characterized by progressive bone loss and destruction, which is usually closely related to diabetes, HIV, etc. However, CN caused by syringomyelia accounts for only 5% of CN cases; the shoulder and elbow are most often involved, and the hip joint is rarely affected. As a rare factor, cervical spondylotic myelopathy (CSM) can be associated with syringomyelia, which is scarcely reported in the literature. Here, we present the first case report to date of CN of the hip caused by syringomyelia secondary to CSM. We describe a 76-year-old male patient who was diagnosed with CSM due to neck pain and weakness of limbs 16 years ago. Four years ago, he noticed recurrent swelling of the right hip with pain and was diagnosed with degenerative arthritis. Recently, however, his symptoms gradually worsened, and because of progressive pain, destabilization and weakness of the right hip, he was admitted to our hospital. Through systematic physical, radiographic and laboratory examinations, we finally reached a diagnosis: CN of the right hip associated with syringomyelia secondary to CSM. After comprehensive evaluation of the patient's condition, we performed right total hip arthroplasty. During the follow-up, the patient felt well clinically and could walk independently with a knee brace. We suggest a possible etiological association between CSM and syringomyelia, which may reflect a potential pathogenesis of CN. We encourage clinicians to actively carry out a detailed medical history and comprehensive physical and imaging examinations in patients with joint lesions, especially chronic shoulder neck pain, to rule out the possibility of this association, which plays a crucial role in the early diagnosis of CN. Arthroplasty may no longer be an absolute contraindication to surgical treatment of CN. Reasonable selection of the surgical strategy can markedly improve the clinical symptoms and quality of life of patients.


Accurate diagnosis of renal transplant rejection by indium-111 platelet imaging despite postoperative cyclosporin therapy.
Previous reports indicate that In-111 platelet scintigraphy (IPS) is a reliable test for the early diagnosis of acute post-operative renal transplant rejection (TR). However, the recent introduction of cyclosporin for post-transplantation immunosuppression requires that the diagnostic efficacy of IPS once again be established. Therefore, a prospective IPS study of 73 post-operative renal transplant recipients was conducted. Fourty-nine patients received cyclosporin and 24 patients did not receive this drug. Between these two patient groups, there were no significant differences in the diagnostic sensitivities (0.86 vs 0.80) and specificities (0.93 vs 0.84) with which TR was identified. We conclude that during the first two weeks following renal transplantation the cyclosporin treatment regimen used at our institution does not limit the reliability of IPS as a test for TR.


Environmental pH affects photoautotrophic growth of Synechocystis sp. PCC 6803 strains carrying mutations in the lumenal proteins of PSII.
Synechocystis sp. strain PCC 6803 grows photoautotrophically across a broad pH range, but wild-type cultures reach a higher density at elevated pH; however, photoheterotrophic growth is similar at high and neutral pH. A number of PSII mutants each lacking at least one lumenal extrinsic protein, and carrying a second PSII lumenal mutation, are able to grow photoautotrophically in BG-11 medium at pH 10.0, but not pH 7.5. We investigated the basis of this pH effect and observed no pH-specific change in variable fluorescence yield from PSII centers of the wild type or the pH-dependent ΔPsbO:ΔPsbU and ΔPsbV:ΔCyanoQ strains; however, 77 K fluorescence emission spectra indicated increased coupling of the phycobilisome (PBS) antenna at pH 10.0 in all mutants. DNA microarray data showed a cell-wide response to transfer from pH 10.0 to pH 7.5, including decreased mRNA levels of a number of oxidative stress-responsive transcripts. We hypothesize that this transcriptional response led to increased tolerance against reactive oxygen species and in particular singlet oxygen. This response enabled photoautotrophic growth of the PSII mutants at pH 10.0. This hypothesis was supported by increased resistance of all strains to rose bengal at pH 10.0 compared with pH 7.5.


Safety and immunogenicity of multiple conventional immunizations administered during early HIV infection.
Twenty-one asymptomatic adults who had recently received multiple polysaccharide, live viral, and protein-derived vaccines were identified as being infected with human immunodeficiency virus (HIV). The mean subject age was 24 years (range 18-33); 20 of 21 (95%) were male. The mean T4 count was 523/mm3 with a mean T4/T8 ratio of 0.6. Serologic responses to immunization with meningococcus group C, adenovirus types 4 and 7, tetanus, and diphtheria were evaluated for the HIV seropositive subjects and were compared with the responses of similarly vaccinated age-, sex-, and race-matched HIV-seronegative controls. Significantly fewer (p less than 0.03) HIV subjects responded to meningococcus C (bactericidal antibody) and adenovirus 4 (neutralizing antibody) vaccines than did normals; the HIV-infected subjects who did respond produced functional antibody comparable to that of normals. Booster responses of HIV subjects to tetanus and diphtheria were comparable to those of normals. HIV-infected vaccine nonresponders did not differ from HIV-infected responders in total white blood cell, T4, T4/T8, total serum IgG, or delayed-type hypersensitivity skin test reactivity. All HIV subjects had negative cultures for live vaccine viruses (rubella, measles, adenovirus, and poliovirus). Postimmunization, no clinically apparent adverse reactions to vaccination were detected.


Comparative histopathological characteristics of highly pathogenic avian influenza (HPAI) in chickens and domestic ducks in 2008 Korea.
We compared characteristic lesions occurring in chickens and domestic ducks naturally infected with H5N1 HPAI virus in April and May 2008. Infected chickens generally exhibited pale-green, watery diarrhoea, depression, neurological signs and cyanosis of wattles and combs, and infected ducks generally exhibited neurological signs and watery diarrhoea. Gross petechial or ecchymotic haemorrhage affected the heart, proventriculus, liver, muscle, fat, and pancreas in chickens, and muscle in ducks. Necrotic foci were primarily present in the pancreas of both species and in the heart of domestic ducks. Histopathologically, chickens exhibited multifocal encephalomalacia, multifocal lymphohistiocytic myocarditis, multifocal necrotic pancreatitis and haemorrhage of several organs and tissues; ducks exhibited lymphohistiocytic meningoencephalitis with multifocal haemorrhages, multifocal necrotic pancreatitis, and severe necrotic myocarditis with mineralisation. The characteristic histopathologic findings of 2008 HPAI were multifocal encephalomalacia and necrotic pancreatitis accompanied by lymphohistiocytic myocarditis, and haemorrhage in various organs and tissues in chickens, whereas in ducks, they were severe necrotic myocarditis with mineralisation and necrotic pancreatitis, accompanied with lymphohistiocytic meningoencephalitis. The high mortality of domestic ducks may be intimately associated with heart failure resulting from increased H5N1 HPAI viral cardiotropism.


High consumption of salt-fermented vegetables and hypertension risk in adults: a 12-year follow-up study.
The aim of this study was to investigate the causal relationship between high consumption of salt-fermented vegetables and hypertension risk in adults. Data came from the Korean Genome and Epidemiology Study, an ongoing community-based cohort study that began in 2001. In the final analysis, a total of 5,932 participants (men=2,822, women=3,110) was included. Daily energy, nutrient, and major salt-fermented vegetables for Korean (kimchi) intakes were assessed using a semi-quantitative food frequency questionnaire. Relative risks and 95% CIs associated with kimchi intake by gender and body mass index (BMI) were estimated using the multivariate Cox proportional hazards regression model. Out of the 5,932 participants, 1,798 (905 men, 893 women) developed hypertension during the 12-year follow-up period. A significant difference in baseline BMI was shown between the non-hypertension and hypertension groups. There was no significant difference with regard to the risk of developing hypertension across quintiles for total kimchi intake and quartile or quartiles for specific kimchi intake in multivariate models by gender and baseline BMI. The trend for increased risk of hypertension according to increasing quartile of watery kimchi intake was significant for obese men in the multivariate model (p<0.05). High consumption of salt-fermented vegetables was not shown to be associated with increased risk of hypertension. The trend for increased risk of hypertension according to increasing quartile of watery kimchi intake was significant only in obese men.


Hepatotoxicity of the combination of rifampin-ethionamide in the treatment of multibacillary leprosy.
During treatment of multibacillary leprosy with the combination rifampin (RMP) 600 mg, ethionamide (ETH) 500 mg, and either dapsone (DDS) or clofazimine (CLO) 100 mg, hepatotoxicity was observed in 4.5% of 596 patients. Hepatitis appeared after 5-186 days, with a mean of 93 days and a median of 76 days. Mortality was 26%. ETH and DDS or CLO were administered daily in all regimens in which hepatitis occurred. RMP was given either daily or daily during the first two weeks or eight weeks, followed by a once-weekly dose. It is concluded that the combination RMP + ETH is the toxic component. In some patient groups there was a high correlation of toxicity with age. A regimen in which RMP was administered only twice a week during three months was not accompanied by hepatotoxicity. Future studies should show if reduction of the daily dose of ETH or reduction of the duration of the administration of RMP + ETH might reduce the incidence of hepatotoxicity while conserving the efficacy.


Seizures and anticonvulsants after aneurysmal subarachnoid hemorrhage.
Seizures and seizure-like activity may occur in patients experiencing aneurysmal subarachnoid hemorrhage. Treatment of these events with prophylactic antiepileptic drugs remains controversial. An electronic literature search was conducted for English language articles describing the incidence and treatment of seizures after aneurysmal subarachnoid hemorrhage from 1980 to October 2010. A total of 56 articles were included in this review. Seizures often occur at the time of initial presentation or aneurysmal rebleeding before aneurysm treatment. Seizures occur in about 2% of patients after invasive aneurysm treatment, with a higher incidence after surgical clipping compared with endovascular repair. Non-convulsive seizures should be considered in patients with poor neurological status or deterioration. Seizure prophylaxis with antiepileptic drugs is controversial, with limited data available for developing recommendations. While antiepileptic drug use has been linked to worse prognosis, studies have evaluated treatment with almost exclusively phenytoin. When prophylaxis is used, 3-day treatment seems to provide similar seizure prevention with better outcome compared with longer-term treatment.


Nomogram for predicting the preoperative lymph node metastasis in resectable pancreatic cancer.
Lymph node metastasis (LNM) is a critical prognostic factor in resectable pancreatic cancer (PC) patients, determining treatment strategies. This study aimed to develop a clinical model to adequately and accurately predict the risk of LNM in PC patients. 13,200 resectable PC patients were enrolled from the SEER (Surveillance, Epidemiology, and End Results) database, and randomly divided into a training group and an internal validation group at a ratio of 7:3. An independent group (n = 62) obtained from The First Affiliated Hospital of Xinxiang Medical University was enrolled as the external validation group. The univariate and multivariate logistic regression analyses were used to screen independent risk factors for LNM. The minimum Akaike's information criterion (AIC) was performed to select the optimal model parameters and construct a nomogram for assessing the risk of LNM. The performance of the nomogram was assessed by the receiver operating characteristics (ROC) curve, calibration plot, and decision curve analysis (DCA). In addition, an online web calculator was designed to assess the risk of LNM. A total of six risk predictors (including age at diagnosis, race, primary site, grade, histology, and T-stage) were identified and included in the nomogram. The areas under the curves (AUCs) [95% confidential interval (CI)] were 0.711 (95%CI: 0.700-0.722), 0.700 (95%CI: 0.683-0.717), and 0.845 (95%CI: 0.749-0.942) in the training, internal validation and external validation groups, respectively. The calibration curves showed satisfied consistency between nomogram-predicted LNM and actual observed LNM. The concordance indexes (C-indexes) in the training, internal, and external validation sets were 0.689, 0.686, and 0.752, respectively. The DCA curves of the nomogram demonstrated good clinical utility. We constructed a nomogram model for predicting LNM in pancreatic cancer patients, which may help oncologists and surgeons to choose more individualized clinical treatment strategies and make better clinical decisions.


[Potential parenthood for the mentally handicapped; the role of the physician].
* What responsibilities and options do physicians have regarding contraception, pregnancy and parenthood in people with mental disability? The issue encompasses the assessment of parenting capability and mental competence, and options for supporting responsible parenthood and preventing irresponsible parenthood. * Parenting capability is a combined trait of both parents. Family and friends may support that capability. * According to a recent description, parenthood is 'good enough' when there are no indications for neglect or abuse, and when there is no involvement of the Child Care and Protection Board. * Mental competence regarding pregnancy signifies that the person involved can understand and weigh the consequences of pregnancy for themselves and others, including the future child. * If parenthood is likely to be irresponsible and the person is sexually active and refuses contraception, a physician has three options: persuasion, pressure or coercion. The latter is legally allowed only in cases of mental incompetence. * People with mental disability will continue to have children, regardless of whether their environment considers it responsible. Although informal caregivers will provide the primary support in this situation, adequate professional support is desirable.


Distribution and number of Cajal-like cells in testis tissue with azoospermia.
We investigated the number and distribution of Cajal-like cells in patients with azoospermia. A total of 99 patients with non-obstructive azoospermia were divided into subgroups [19 patientsin hypospermatogenesis group (S1), 40 patients in maturation arrest group (S2), 20 patients in a Sertoli cell-only syndrome (S3), and 20 patients in a testicular atrophy and fibrosis group (S4)], and 20 patients with obstructive azoospermia group (S0). Sections stained with a c-kit antibody were studied by light microscopy to determine the number and distribution of Cajal-like cells in peritubular and perivascular areas of testis. The number of Cajal-like cells were higher in all the non-obstructive groups than in the obstructive group (S0: 2.43 cells/mm2, S1: 3.14 cells/mm2, S2: 4.00 cells/mm2, S3: 4.57 cells/mm2, S4: 3.86 cells/mm2) but statistically significantly different (p<0.05) in the S2 and S3 subgroups only. Distribution of Cajal-like cells were similar in all groups. The number and distribution of Cajal-like cells in non-obstructive groups suggest that these cells may affect spermatogenesis. This cellular type can be responsible for the regulation of cellular motility or spermatogenesis. Electrophysiological and electron microscopic studies are needed to better define morphology and function of Cajal-like cells in the testis, especially totally the normal testis tissue.


Targeted Delivery of Recombinant Heat Shock Protein 27 to Cardiomyocytes Promotes Recovery from Myocardial Infarction.
Ischemic heart disease, especially myocardial infarction (MI), is the leading cause of death worldwide. Apoptotic mechanisms are thought to play a significant role in cardiomyocyte death after MI. Increased production of heat shock proteins (Hsps) in cardiomyocytes is a normal response to promote tolerance and to reduce cell damage. Hsp27 is considered to be a therapeutic option for the treatment of ischemic heart disease due to its protective effects on hypoxia-induced apoptosis. Despite its antiapoptotic effects, the lack of strategies to deliver Hsp27 to the heart tissue in vivo limits its clinical applicability. In this study, we utilized an antibody against the angiotensin II type 1 (AT1) receptor, which is expressed immediately after ischemia/reperfusion in the heart of MI rats. To achieve cardiomyocyte-targeted Hsp27 delivery after ischemia/reperfusion, we employed the immunoglobulin-binding dimer ZZ, a modified domain of protein A, in conjunction with the AT1 receptor antibody. Using the AT1 receptor antibody, we achieved systemic delivery of ZZ-TAT-GFP fusion protein into the heart of MI rats. This approach enabled selective delivery of Hsp27 to cardiomyocytes, rescued cells from apoptosis, reduced the area of fibrosis, and improved cardiac function in the rat MI model, thus suggesting its applicability as a cardiomyocyte-targeted protein delivery system to inhibit apoptosis induced by ischemic injury.


Antifungal activities of three different Lactobacillus species and their production of antifungal carboxylic acids in wheat sourdough.
This study was undertaken to assess the antifungal performance of three different Lactobacillus species.Experiments were conducted in vitro and in situ to extend the shelf life of wheat bread. Standard sourdough analyses were performed characterising acidity and carbohydrate levels. Overall, the strains showed good inhibition in vitro against the indicator mould Fusarium culmorum TMW4.2043. Sourdough bread fermented with Lactobacillus amylovorus DSM19280 performed best in the in situ shelf life experiment. An average shelf life extension of six more mould-free days was reached when compared to the non-acidified control bread. A range of antifungal-active acids like 3-phenyllactic acid, 4-hydroxyphenyllactic acid and 2-hydroxyisocaproic acid in quantities between 0.1 and 360 mg/kg were present in the freeze-dried sourdoughs. Their concentration differed greatly amongst the species.However, a higher concentration of these compounds could not completely justify the growth inhibition of environmental moulds. In particular, although Lb. reuteri R29 produced the highest total concentration of these active compounds in the sourdough, its addition to bread did not result in a longest shelf life. Nevertheless, when the artificial compounds were spiked into a chemically acidified dough, it succeeded in a longer shelf life (+25 %) than achieved only by acidifying the dough. This provides evidence of their contribution to the antifungal activity and their synergy in concentration levels far below their single minimal inhibition concentrations under acidic conditions.


Sox2 is required for embryonic development of the ventral telencephalon through the activation of the ventral determinants Nkx2.1 and Shh.
The Sox2 transcription factor is active in stem/progenitor cells throughout the developing vertebrate central nervous system. However, its conditional deletion at E12.5 in mouse causes few brain developmental problems, with the exception of the postnatal loss of the hippocampal radial glia stem cells and the dentate gyrus. We deleted Sox2 at E9.5 in the telencephalon, using a Bf1-Cre transgene. We observed embryonic brain defects that were particularly severe in the ventral, as opposed to the dorsal, telencephalon. Important tissue loss, including the medial ganglionic eminence (MGE), was detected at E12.5, causing the subsequent impairment of MGE-derived neurons. The defect was preceded by loss of expression of the essential ventral determinants Nkx2.1 and Shh, and accompanied by ventral spread of dorsal markers. This phenotype is reminiscent of that of mice mutant for the transcription factor Nkx2.1 or for the Shh receptor Smo. Nkx2.1 is known to mediate the initial activation of ventral telencephalic Shh expression. A partial rescue of the normal phenotype at E14.5 was obtained by administration of a Shh agonist. Experiments in Medaka fish indicate that expression of Nkx2.1 is regulated by Sox2 in this species also. We propose that Sox2 contributes to Nkx2.1 expression in early mouse development, thus participating in the region-specific activation of Shh, thereby mediating ventral telencephalic patterning induction.


Microscale 3D collagen cell culture assays in conventional flat-bottom 384-well plates.
Three-dimensional (3D) culture systems such as cell-laden hydrogels are superior to standard two-dimensional (2D) monolayer cultures for many drug-screening applications. However, their adoption into high-throughput screening (HTS) has been lagging, in part because of the difficulty of incorporating these culture formats into existing robotic liquid handling and imaging infrastructures. Dispensing cell-laden prepolymer solutions into 2D well plates is a potential solution but typically requires large volumes of reagents to avoid evaporation during polymerization, which (1) increases costs, (2) makes drug penetration variable and (3) complicates imaging. Here we describe a technique to efficiently produce 3D microgels using automated liquid-handling systems and standard, nonpatterned, flat-bottomed, 384-well plates. Sub-millimeter-diameter, cell-laden collagen gels are deposited on the bottom of a ~2.5 mm diameter microwell with no concerns about evaporation or meniscus effects at the edges of wells, using aqueous two-phase system patterning. The microscale cell-laden collagen-gel constructs are readily imaged and readily penetrated by drugs. The cytotoxicity of chemotherapeutics was monitored by bioluminescence and demonstrated that 3D cultures confer chemoresistance as compared with similar 2D cultures. Hence, these data demonstrate the importance of culturing cells in 3D to obtain realistic cellular responses. Overall, this system provides a simple and inexpensive method for integrating 3D culture capability into existing HTS infrastructure.


Statins markedly potentiate aminopeptidase inhibitor activity against (drug-resistant) human acute myeloid leukemia cells.
Aim: This study aimed to decipher the molecular mechanism underlying the synergistic effect of inhibitors of the mevalonate-cholesterol pathway (i.e., statins) and aminopeptidase inhibitors (APis) on APi-sensitive and -resistant acute myeloid leukemia (AML) cells. Methods: U937 cells and their sublines with low and high levels of acquired resistance to (6S)-[(R)-2-((S)-Hydroxy-hydroxycarbamoyl-methoxy-methyl)-4-methyl-pentanoylamino]-3,3 dimethyl-butyric acid cyclopentyl ester (CHR2863), an APi prodrug, served as main AML cell line models. Drug combination effects were assessed with CHR2863 and in vitro non-toxic concentrations of various statins upon cell growth inhibition, cell cycle effects, and apoptosis induction. Mechanistic studies involved analysis of Rheb prenylation required for mTOR activation. Results: A strong synergy of CHR2863 with the statins simvastatin, fluvastatin, lovastatin, and pravastatin was demonstrated in U937 cells and two CHR2863-resistant sublines. This potent synergy between simvastatin and CHR2863 was also observed with a series of other human AML cell lines (e.g., THP1, MV4-11, and KG1), but not with acute lymphocytic leukemia or multiple solid tumor cell lines. This synergistic activity was: (i) specific for APis (e.g., CHR2863 and Bestatin), rather than for other cytotoxic agents; and (ii) corroborated by enhanced induction of apoptosis and cell cycle arrest which increased the sub-G1 fraction. Consistently, statin potentiation of CHR2863 activity was abrogated by co-administration of mevalonate and/or farnesyl pyrophosphate, suggesting the involvement of protein prenylation; this was experimentally confirmed by impaired Rheb prenylation by simvastatin. Conclusion: These novel findings suggest that the combined inhibitory effect of impaired Rheb prenylation and CHR2863-dependent mTOR inhibition instigates a potent synergistic inhibition of statins and APis on human AML cells.


Endoscopic removal of a right lower lobe bronchial lipoma in a high-risk patient: a case report.
Endobronchial lipomas are rare benign tumours that cause bronchial obstruction, leading to extensive distal parenchymal damage. This report describes the endoscopic removal of a right main bronchus lipoma with a rigid endoscope and snare technique in a high-risk patient. The patient was successfully treated without complication. Endoscopic reaction is the preferred therapeutic approach for this subgroup of patients.


Tectorigenin, an isoflavone of Pueraria thunbergiana Benth., induces differentiation and apoptosis in human promyelocytic leukemia HL-60 cells.
Cytotoxic effects of six isoflavonoids, tectorigenin, glycitein, tectoridin, glycitin, 6''-O-xylosyltectoridin, and 6''-O-xylosylglycitin isolated from the flower of Pueraria thunbergiana Benth. together with genistein, a known differentiation and apoptosis inducer, were examined. Among these isoflavonoids, tectorigenin and genistein exhibited cytotoxicity against various human cancer cells; glycitein showed only mild cytotoxicity. These results suggest that the isoflavone structure and 5-hydroxyl group are crucial for the cytotoxic properties and that glycosides are inactive. Moreover, tectorigenin induced differentiation of human promyelocytic leukemia HL-60 cells to granulocytes and monocytes/macrophages, and caused apoptotic changes of DNA in the cells, as did genistein. Tectorigenin also inhibited autophosphorylation of epidermal growth factor (EGF) receptor by EGF and decreased the expression of Bcl-2 protein, with less activity than genistein. From these results, tectorigenin may be a possible therapeutic agent for leukemia.


Fasting blood glucose as a predictor of progressive infarction in men with acute ischemic stroke.
Blood glucose is related to early neurological deterioration in acute ischemic stroke, but multiple mechanisms are involved in early neurological deterioration, such as progressive infarction. This study aimed to determine whether fasting blood glucose (FBG) is an independent predictor of progressive infarction. From April 2017 to December 2020, we retrospectively enrolled 477 patients with acute ischemic stroke within 48 hours of onset. Demographic characteristics, clinical information, neuroimaging characteristics, and laboratory data were collected after admission. We found that 147 (30.8%) patients had progressive infarction. Multiple regression analysis showed that high FBG concentrations (>7.66 mmol/L) were independently associated with progressive infarction. Sex subgroup analysis showed that high FBG concentrations were an independent predictor of progressive infarction in male patients (odds ratio, 2.559; 95% confidence interval, 1.279-5.121). In a receiver operating characteristic curve analysis, FBG concentrations were a predictor of progressive infarction in all cases, especially in male patients. The cutoff value of FBG in all patients and men was 7.155 mmol/L. FBG is an independent predictor of progressive infarction in patients with acute ischemic stroke within 48 hours of onset, especially in men. Patients with FBG concentrations ≥7.155 mmol/L are more likely to develop progressive infarction.


Spontaneous spinal epidural haematoma due to arteriovenous malformation in a child.
Spontaneous spinal epidural haematoma (SSEH) is a rare clinical entity, especially in infants, in whom only a few cases have been reported. In a paediatric emergency setting, SSEH should be considered as part of the differential diagnosis for acute extremity weakness and paraesthesia. Epidural vascular malformations are often suspected in these cases but have rarely been demonstrated. The authors report herein a case of SSEH in a 9-year-old boy arising from an epidural vascular malformation. He initially presented with sudden intense cervicodorsal pain followed by hypotonic lower extremities and progressive motor weakness, with no sensory change. The MRI showed an acute extradural haematoma extending from C7 to T4 with compression of the spinal cord. After submission to decompression surgery, he presented full recovery in 1 month. The histopathological analysis revealed a vascular malformation.


Constructing honeycomb micropatterns on nonplanar substrates with high glass transition temperature polymers.
In Qiao's previous report, only star polymers with T(g) (glass transition temperature) below 48°C were found forming homogeneous honeycomb coatings on the nonplanar substrates. The polymers with high T(g) are believed not able to duplicate nonplanar substrate due to their brittleness. This article presents a comprehensive study on the construction of macroporous polymeric films on various nonplanar substrates with static breath figure (BF) technique, using linear polymers with high T(g). Two kinds of linear polymers with high T(g), polystyrene-b-poly(acrylic acid) and polystyrene without polar end groups, are employed to prepare 3-dimensional macroporous films on different nonplanar substrates. Scanning electronic microscopy views on the side wall in addition to views in-plane prove that polymer films with BF array perfectly replicated the surface features of these substrates. The formation processes of macropores on these substrates are analyzed in detail, and it demonstrates that neither molecular topography nor T(g) of polymers is the critical factor contouring nonplanar substrate. A new hypothesis involving polymer plasticization and conformation during the solvent evaporation is formulated.


Evaluation of proinflammatory cytokines and inflammation markers as biomarkers for the action of thiazolidinediones in Type 2 diabetes mellitus patients and healthy volunteers.
Thiazolidinediones (TZDs) not only enhance cellular glucose transport but are reported to have potent anti-inflammatory effects. These effects may play an important role in the insulin sensitizing mechanism, and possibly precede the effects on parameters of glucoregulation. We sought to investigate whether these anti-inflammatory effects could yield early responding biomarkers for TZD action in Type 2 diabetes mellitus (T2DM) patients and healthy volunteers (HV) to expedite early clinical development of novel compounds. We investigated the timing of treatment effects on several proinflammatory cytokines and markers of inflammation in comparison with effects on typical measures of glucoregulation in T2DM patients and HV receiving rosiglitazone 4 mg or placebo twice daily for 6 weeks. We found a significant reduction in interleukin (IL)-6 [-39.4%, confidence interval (CI) - 60.0, - 8.2] and white blood cell count (-18.4%, CI - 30.2, - 4.5) after 4 weeks of treatment in the T2DM group. These anti-inflammatory effects did not precede the effects on typical parameters of glucoregulation in the T2DM group and there was no significant anti-inflammatory response in the HV group. We could not identify biomarkers that precede the effects of rosiglitazone on parameters of glucoregulation in T2DM or that have a significant response in HV. However, the IL-6 response observed in this study indicates a potential role for this cytokine as complementary biomarker in clinical 'proof of concept' studies with novel TZDs.


Measurements of buccal tissue volumes at single-implant restorations after local bone grafting in maxillas: a 3-year clinical prospective study case series.
The purpose of this study was to measure changes in buccal and proximal tissue volumes after local bone grafting and single-implant treatment. Ten patients were provided with buccal bone grafts 6 months prior to implant treatment in central upper incisor regions. Following a healing time of 6 months, abutments and single-implant crowns were installed and followed up for 2 years. Clinical photographs and impressions were taken prior to the surgical intervention as well as after crown placement and at first and second annual checkups. The photographs and study models were analyzed with regard to papilla regeneration and changes in buccal crest volume during the study period by means of a clinical papilla index and optical scanning of study models. All bone grafts healed without problems. A significant reduction of the buccal crest volume (-50%, p <.01) was observed in the grafted area before abutment connection. However, a significant increase of tissue volume (+100%, p <.05) was noticed at the subsequent crown placement, followed by a second but slow reduction of the volume during the following 2 years of function. The interdental papillae increased significantly (p <.05) in volume during the first year, almost completely filling up the embrasure areas after 2 years. It may be concluded that local bone grafting seems to be a valuable protocol to create sufficient bone volume for implant placement. However, significant resorption of the graft may be present, which reduces the impact of grafting on the esthetic outcome. Instead, placement of the abutment cylinder and the crown seems to play a more important role for reestablishing the tissue volume at the implant-supported single crowns.


Heterogeneity of mast cells and expression of Annexin A1 protein in a second degree burn model with silver sulfadiazine treatment.
Mast cells (MCs) participate in all stages of skin healing and one of their mediators is the Annexin A1 protein (AnxA1), linked to inflammation, proliferation, migration and apoptosis processes, but not studied in thermal burns yet. Therefore, our objectives were to evaluate the behavior of MCs and AnxA1 in a second degree burn model, treated or not with silver sulfadiazine 1% (SDP 1%) and associated to macrophages quantification and cytokines dosages. MCs counts showed few cells in the early stages of repair but increased MCs in the final phases in the untreated group. The normal skin presented numerous tryptase-positive MCs that were reduced after burning in all analyzed periods. Differently, few chymase-positive MCs were observed in the early stages of healing, however, increased chymase-positive MCs were found at the final phase in the untreated group. MCs also showed high immunoreactivity for AnxA1 on day 3 in both groups. In the tissue there was a strong protein expression in the early stages of healing, but in the final phases only in the SDP treated animals. TNF-α, IL-1β, IL-6, IL-10 and MCP-1 levels and macrophages quantification were increased in inflammation and reepithelialization phases. Reduced IL-1β, IL-6 and IL-10 levels and numerous macrophages occurred in the treated animals during tissue repair. Our results indicate modulation in the profile of MCs and AnxA1expression during healing by the treatment with SDP 1%, pointing them as targets for therapeutic interventions on skin burns.


Stream macroinvertebrate drift response to pulsed exposure of the synthetic pyrethroid lambda-cyhalothrin.
Outdoor experimental channels were used to study the behavioral changes of stream macroinvertebrates exposed to a pulse of the pyrethroid insecticide lambda-cyhalothrin. The primary end point was the number of macroinvertebrates drifting, but the mobility of macroinvertebrates caught in the drift also was assessed. A specified number of two insect species, Baetis rhodani and Leuctra fusca/digitata, and of the amphipod Gammarus pulex were introduced into small replicated subsections of the experimental channels. Macroinvertebrates were allowed to acclimatize for 26 h prior to a 60-min pulsed exposure to lambda-cyhalothrin. Measurement was initiated 2 h before pesticide application and continued for the following 24 h. Pulse concentrations of lambda-cyhalothrin of 0.001, 0.01, 0.1, and 1.0 microg L(-1) were applied, and each experiment was replicated 8 times. All three species responded to the pesticide pulse with catastrophic drift. The 0.001 microg L(-1) treatment caused a significant increase in the drift of Gammarus, whereas the drift response threshold was 0.01 microg L(-1) for the two insect species. Drift response onset followed the applied pulse concentration, with the highest concentrations resulting in more individuals of all species entering drift at an early stage. The majority of individuals caught in drift samples during low concentrations showed no change in mobility. At the two highest concentrations, however, both Baetis and Leuctra were in the process of being immobilized, with Leuctra the more sensitive of the two. In contrast, only a few of the Gammarus individuals caught showed changes in mobility after the high-concentration treatments. The present study shows that lambda-cyhalothrin is a potential hazard for macroinvertebrate populations in headwater streams. The clear species-specific responses indicate that sublethal doses have the potential to change the macroinvertebrate community structure.


Genetic determinants of neoplastic diseases induced by a subgroup F avian leukosis virus.
Two subgroup F avian leukosis viruses, ring-necked pheasant virus (RPV) and RAV-61, were previously shown to induce a high incidence of a fatal proliferative disorder in the lungs of infected chickens. These lung lesions, termed angiosarcomas, appear rapidly (4 to 5 weeks after infection), show no evidence of proto-oncogene activation by proviral integration, and are not induced by avian leukosis viruses belonging to other subgroups. To identify the viral sequences responsible for induction of these tumors, we constructed recombinant viruses by exchanging genomic segments of molecularly cloned RPV with those of a subgroup A leukosis virus, UR2AV. The ability to induce rapid lung tumors segregated only with the env sequences of RPV; the long terminal repeat of RPV was not required. However, recombinants carrying both env and long terminal repeat sequences of RPV induced lung tumors with a shorter latency. In several cases, recombinant viruses exhibited pathogenic properties differing from those of either parental virus. Recombinants carrying the gag-pol region of RPV and the env gene of UR2AV induced a high incidence of a muscle lesion termed infiltrative intramuscular fibromatosis. One recombinant, EU-8, which carries the gag-pol and LTR sequences of RPV, and the env gene of UR2AV, induced lymphoid leukosis after an unusually short latent period. The median time of death from lymphoid leukosis was 6 to 7 weeks after infection with EU-8 compared with approximately 5 months for UR2AV.


Combined effects of Chinese medicine feed and ginger extract bath on co-infection of Ichthyophthirius multifiliis and Dactylogyrus ctenopharyngodonid in grass carp.
Dactylogyrus ctenopharyngodonid and Ichthyophthirius multifiliis are two important ectoparasites of freshwater fish. Co-infection by the two parasites leads to high fish mortality and results in heavy economic losses. This study aimed to evaluate the efficacy of medicated feed and a ginger extract bath against D. ctenopharyngodonid and I. multifiliis on grass carp and investigate the hematological response of grass carp co-infected by the two parasites. These results demonstrated that red blood cell (RBC) and thrombocyte percentage among leucocytes significantly decreased after grass carp were co-infected by D. ctenopharyngodonid and I. multifiliis. The monocyte and neutrophil percentages significantly increased with the increment of parasite mean intensities, while the lymphocyte percentage decreased. The activities of serum acid phosphatase (ACP), alkaline phosphatase (AKP), lysozyme (LZM), and superoxide dismutase (SOD) significantly increased after co-infection. When grass carp treated with medicated feed containing 4% of Astragalus membranaceus, Allium sativum, Morus alba, and Glycyrrhiza uralensis, the activities of ACP, AKP, LZM, and SOD were significantly enhanced, and the mean intensities of D. ctenopharyngodonid and I. multifiliis were significantly decreased. When grass carp was treated with medicated feed and a 4-mg/L ginger extract bath, all parasites were eliminated during 28 days. The bath of ginger extract at a concentration of 4 mg/L kept a low mean intensity of I. multifiliis and D. ctenopharyngodonid, then the two parasites were eliminated by oral administration of the medicated feed with an immunostimulant (Chinese medicine compound).


Plant-derived H7 VLP vaccine elicits protective immune response against H7N9 influenza virus in mice and ferrets.
In March 2013, the Chinese Centre for Disease Control and Prevention confirmed the first reported case of human infection with an avian influenza A H7N9 virus. Infection with this virus often caused severe pneumonia and acute respiratory distress syndrome resulting in a case fatality rate >35%. The risk of pandemic highlighted, once again, the need for a more rapid and scalable vaccine response capability. Here, we describe the rapid (19 days) development of a plant-derived VLP vaccine based on the hemagglutinin sequence of influenza H7N9 A/Hangzhou/1/2013. The immunogenicity of the H7 VLP vaccine was assessed in mice and ferrets after one or two intramuscular dose(s) with and without adjuvant (alum or GLA-SE™). In ferrets, we also measured H7-specific cell-mediated immunity. The mice and ferrets were then challenged with H7N9 A/Anhui/1/2013 influenza virus. A single immunization with the adjuvanted vaccine elicited a strong humoral response and protected mice against an otherwise lethal challenge. Two doses of unadjuvanted vaccine significantly increased humoral response and resulted in 100% protection with significant reduction of clinical signs leading to nearly asymptomatic infections. In ferrets, a single immunization with the alum-adjuvanted H7 VLP vaccine induced strong humoral and CMI responses with antigen-specific activation of CD3(+) T cells. Compared to animals injected with placebo, ferrets vaccinated with alum-adjuvanted vaccine displayed no weight loss during the challenge. Moreover, the vaccination significantly reduced the viral load in lungs and nasal washes 3 days after the infection. This candidate plant-made H7 vaccine therefore induced protective responses after either one adjuvanted or two unadjuvanted doses. Studies are currently ongoing to better characterize the immune response elicited by the plant-derived VLP vaccines. Regardless, these data are very promising for the rapid production of an immunogenic and protective vaccine against this potentially pandemic virus.


Acetate Production from Anaerobic Oxidation of Methane via Intracellular Storage Compounds.
There is great interest in microbial conversion of methane, an abundant resource, into valuable liquid chemicals. While aerobic bioconversion of methane to liquid chemicals has been reported, studies of anaerobic methane bioconversion to liquid chemicals are rare. Here we show that a microbial culture dominated by Candidatus 'Methanoperedens nitroreducens', an anaerobic methanotrophic archaeon, anaerobically oxidizes methane to produce acetate, indirectly via reaction intermediate(s), when nitrate or nitrite is supplied as an electron acceptor under a rate-limiting condition. Isotopic labeling tests showed that acetate was produced from certain intracellular storage compounds that originated from methane. Fluorescence in situ hybridization and Nile red staining demonstrated that polyhydroxyalkanoate in M. nitroreducens was likely one of the intracellular storage compounds for acetate production, along with glycogen. Acetate is a common substrate for the production of more valuable chemicals. The microbial conversion discovered in this study potentially enables a new approach to the use of methane as a feedstock for the chemical market.